Monoclonal antibodies have wide applications in diagnosis and treatment. They can be used as diagnostic tools in assays and imaging to detect diseases. As therapeutic agents, they can directly target pathogens or cancer cells, or act as drug delivery systems. Regulations require approval from agencies like RCGM, GEAC and CDSCO for research, clinical trials and marketing of monoclonal antibodies and similar biologics in India. Advancements continue to improve antibody design for enhanced safety and efficacy in clinical applications.

1. Applications of
Monoclonal antibody
Dr Saurav Misra
PGIMS Rohtak

2. 1. Diagnostic Application
(A) MAbs in Biochemical Analysis:
Diagnostic tests based on the use of MAbs as reagents are used in
radioimmunoassay (RIA) and enzyme linked immunosorbent assay
Measure the circulating concentrations of hormones ( Insulin,
H.C.G, growth hormone, progesterone)
Measure the circulating concentration of blood products like antigen,
clotting factors and interleukins.

3. Number of diagnostic kits using MAbs have
become available.
It is now possible to do the early diagnosis of the following
conditions/diseases.
1. Pregnancy.
2. Cancer.
3. Hormonal disorder.
4. Infectious disease.

4. B. USES IN DIAGNOSTIC IMAGING:
Radiolabelled MAbs are used in the diagnostic imaging of disease
,this technique is referred to as immune scintigraphy.
Radioisotopes: Iodine 131,Technetium 99
Single photon emission computed tomography (SPECT) cameras
are used
E.g. Nofetumomab, monoclonal antibody that when tagged with the
radioisotope can detect a protein found on the surface of small lung
cancer cells, Capromab pendetide

5. Monoclonal antibodies are successfully used in the diagnostic
imaging of:
1. Cardiovascular diseases
2. Cancers
3. Sites of bacterial infections.

6. A2. Therapeutic Applications
(A) MAbs as Direct Therapeutic Agents:
1. In destroying disease causing organism:
Mabs promote efficient opsonisation of pathogenic organisms and
enhance phagocytosis.
Eg palivizumab, foravirimab, regavirumab, sevirumab
2. Treatment of cancer:
Leukemia, colorectal cancer, lymphoma and melanoma
Eg rituximab, trastuzumab, cituzimab, bevacizumab

7. 3. Immunosuppression in organ transplantation:
MAbs specific to T-lymphocytes surface antigens are used
Eg. OKT (first MAb to be licensed by U.S), alemtuzumab
4. Treatment of AIDS:
Eg: Ibalizumab, Anti CD-4 antibody
5. Treatment of autoimmune diseases:
Eg Etarnacept, Adalimumab, Infliximab

8. (B) MAbs in use as immunotoxins:
Toxins can be coupled with MAbs to form immunotoxins and used in
therapy eg diphtheria toxins, pseudomonas exotoxin
Eg. Anti-Tac MAb raised against IL2-R (T-cell growth factor
receptor) can be conjugated with exotoxin of Pseudomonas sp.
This immunotoxin can be used to destroy the malignant T-cells in
the patients suffering from T-cell leukemia

9. 2)Antibody-directed enzyme prodrug therapy (ADEPT)
➢ In the treatment of certain diseases, a pro-drug (an inactive form of
the drug) can be used.
➢ This can be enzymatically converted to active drug in the target
tissues. For this purpose, the enzyme (that converts pro-drug to
drug) is coupled with MAb that is directed against a specific cell
surface antigen
➢ Eg. Lactamase for hydrolyzing β-lactam ring containing antibiotics.

10. (E) Drug delivery through liposomes coupled to tissue specific MAbs
.
Liposomes are sacs or vesicles formed spontaneously when certain
lipid molecules are exposed to aqueous environment.
Drug entrapped in liposomes are coated with MAbs directed
against tissue specific antigens are being tried for drug delivery.
Eg Anti HER2 antibody + Liposomal Doxorubicin for Ca Breast

11. (F). MAbs in radio immunotherapy (RAIT):
The radioisotopes can be coupled to MAbs that are directed
against tumour cells.
This allows the concentration of radioactivity at the desired sites
and a very efficient killing of target cells (tumor cells).
Eg: Tositumomab used for nonHodgkins lymphoma.

12. 3: Protein purification
Monoclonal antibodies can
also be used to purify a
substance with techniques
called immunoprecipitation
and affinity
chromatography.

13. 4. Miscellaneous Applications:
(A) Catalytic MAbs (ABZYMES):
Mabs incorporating metal ions have been developed to carry out
catalysis.
Abzymes represent a major biotechnological advancement that
have a wide range of applications (cutting of peptides and DNAs,
dissolution of blood clots , killing of viruses)
Eg Anti CEA + Carboxypeptidase G2 For Chorio CA, Ca Breast.

14. (B) Autoantibody Fingerprinting:
IS(individual specific) -autoantibodies are produced after birth and
reach maximum in number by 2 years, and then remain constant for
the later part of life.
Monoclonal antibodies produced against IS (individual specific)
autoantibodies can be used for their detection and identification of
individuals
This technique referred to as autoantibody finger printing is
particularly useful for the detection of criminals and rapists.
The autoantibodies collected from blood, saliva, semen and tears.

15. Antineoplastic
Monoclonal Antibodies

16. Drug Origin
Target
Indication ADRs
1. Trastuzumab Humanized
MAbs
HER2
Ca Breast (
Patients with
HER2/neu-
amplified tumors)
flu-like symptoms
(such as fever, chills
and mild pain),
nausea and diarrhea.
2. Cetuximab Chimeric
(Human,
Mouse)
EGF
Receptor
Head and neck
cancer, colorectal
cancer, lung
cancer
Acneform Rash,
Diarrhea
Cardio-pulmonary
Arrest.

17. Drug Source
Target
Indications ADRs
4. Bevacizumab Humanized
VEGF- A
Lung, renal,
ovarian
cancer,
glioblastoma
multiforme,
breast cancer
Infusion reaction
Bleeding, HTN,
GI Perforation
stroke, MI,
Nephrotic
Syndrome
3. Rituximab Chimeric
Anti CD-20
Lymphoma,
leukemias,
transplant
rejection,
autoimmune
disorder
Infusion reaction ,
cardiac arrest,
cytokine release
syndrome, tumour
lysis syndrome

18. Drug Origin and
target
Indications ADRs
5.
Panitumumab
humanized
IgG2 against
EGFR
Metastatic
colorectal
carcinoma
Rash and dermatological
toxicity
Severe infusion reactions
Pulmonary fibrosis
6.
Alemtuzumab
Humanized
IgG,Against
CD52
CLL , B & T
cell low
grade
lymphomas
Acute infusion reaction
Myelosuppression
Risk of fungal, viral
infections
7. Ipilimumab Human IgG
Binds CTLA-4
(cytotoxic T
lymphocyte-
associated
antigen 4)
Metastatic
Melanoma
T Cell mediated
inflammatory reactions
(Colitis, Hepatitis,
Dermatitis )
GI disturbance diarrhea

19. Anti TNF Agents
Monoclonal Antibodies

20. Drug Source Indications
1. Infliximab Human-mouse
chimeric IgG1
Crohn's disease,
Ulcerative colitis,
Rheumatoid arthritis,
Ankylosing spondylitis,
and
Psoriatic arthritis
Adalimumab Human IgG1 Rheumatoid arthritis
Ankylosing spondylitis
Crohn's disease
juvenile idiopathic
arthritis
plaque psoriasis
psoriatic arthritis

21. Drug Source and target Indications
Certolizumab
pegol
Humanized
FAB fragment
Anti TNF
Crohn's disease ,
Rheumatoid arthritis
Golimumab Human
Anti TNF
moderately to severely active
rheumatoid arthritis, psoriatic
arthritis, and ankylosing
spondylitis
Etarnacept Human anti TNF Rheumatoid arthritis,
juvenile rheumatoid arthritis
and psoriatic arthritis, plaque
psoriasis and ankylosing
spondylitis.

22. Toxicities of Anti TNF agents
Increased risk of serious infections. (tuberculosis)
Increase the risk of lymphoma and possibly other
malignancies.
Can also induce the development of anti-DNA antibodies
Infusion or injection site reactions
Demyelinating central nervous system disease.

23. Monoclonal Antibodies
used in
Autoimmune Diseases

24. Eculizumab
Humanized IgG
Binds the C5 complement component, inhibiting its cleavage into C5a and
C5b thereby inhibiting the terminal pore-forming lytic activity of complement.
Use : Paroxysmal nocturnal hemoglobinuria (PNH)
ADRs: Increased risk of meningococcal infection

25. Natalizumab :
• Humanized monoclonal antibody against α4-integrin (also
known as VLA-4).
ADRs
Increased risk of progressive multifocal leukoencephalopathy (PML)
Uses :
1. Crohn's disease.
2. Multiple Sclerosis

26. Drug Type Target Approved Use
Tocilizumab humanized
monoclonal
antibody
against the
interleukin-6
receptor
(IL-6R)
Castleman's disease
Rheumatoid arthritis
Belimumab Human BLyS (or B-
lymphocyte
stimulator)
SLE
Rheumatoid Arthritis
Basiliximab chimeric
mouse-
human
monoclonal
antibody
α chain
(CD25) of the
IL-2 receptor
prevent rejection in
organ transplantation,
especially in kidney
transplants

27. Drug Type and target Approved use
Daclizumab Humanized,alpha
subunit of the IL-2
receptor of T cells
prevent rejection in
organ transplantation,
especially in kidney
transplants
Ustekinumab Humanized,
against interleukin 12
and interleukin 23
severe plaque
psoriasis
multiple sclerosis and
sarcoidosis

28. Monoclonal Antibodies used in
Infections

29. Drug Possible use
1. Efungumab (Fungus) Invasive Candida infection in
combination with amphotericin B
2. Exbivirumab Hepatitis
3. Foravirimab Prophylaxis of Rabies
4. Libivirumab Hepatitis B
5. Rafivirumab prophylaxis of rabies
6. Regavirumab Infections with cytomegalovirus
7. Sevirumab Infections with cytomegalovirus in
patients with AIDS
8. Tuvirumab Hepatitis B
9. Felvizumab Respiratory syncytial virus

30. Radio-Immuno
conjugates

31. Drug Type
Target
Radioisotope Use
Arcitumomab murine F(ab')
fragment
anti-CEA
antibody
technetium
99m (99mTc)
Imaging patients with
metastatic colorectal
carcinoma
(immunoscintigraphy)
Ibritumomab
tiuxetan
murine
anti-CD20
isotopic
yttrium (90Y)
or 111In
Follicular, or
B-cell non-Hodgkin's
lymphoma
Tositomomab Murine
anti-D20
iodine 131
(131I)
CD20-positive,
follicular non-
Hodgkin's lymphoma

32. Capromab
Pendetide
murine
Anti prostate
specific
membrane
antigen
indium
(111In)
Immunoscintigraphy
for patients with
biopsy-confirmed
prostate cancer
Nofetumomab Murine
40 KD tumor
protein
99m Tc
To stage patients
with small cell lung
cancer
fanolesomab Murine
CD15
technetium-
99m
(99mTc)
Diagnosis of
appendicitis

33. Miscellaneous MABs

34. Abciximab
Fab fragment of a humanized
monoclonal antibody directed against
the II B receptor.
Use:
Patients undergoing percutaneous
angioplasty for coronary thromboses
Adverse Effects:
Bleeding (GI Bleed)
Thrombocytopenia .

35. Omalizumab
Humanized MAB against IgE
administered by subcutaneous
injection every 2-4 weeks
Clinical Use
Asthma prophylaxis
Allergic rhinitis
protection against anaphylaxis
during specific immunotherapy
ADRs:
anaphylactic response, which is
uncommon (<0.1%)

36. Denosumab
Human antibody,
binds with RANKL,
Denosumab blocks osteoclast
formation and activation.
It increases BMD and decreases bone
turnover markers
Use: Osteoporosis

37. Road ahead ….

38. Armed Antibodies
✓ Radioimmunoconjugates
✓ ADEPT
✓ Immunotoxin
✓ Immunocytokine
✓ Immunoliposome
✓ Cellular
Immunoconjugate
✓ Bispescific MABs

39. Single-domain antibody
Consists of a single monomeric variable antibody
domain(VH)
Relatively low molecular wt.(12-15 kDa Vs 120-
150kDa)
better permeability in tissues
they do not show complement system triggered
cytotoxicity because they lack an Fc region
oral administration.
ALX-0081 : Is a single-domain antibody targeting
von Willebrand factor is in clinical trials for the
prevention of thrombosis in patients with acute
coronary syndrome

40. Bi-specific T-cell engagers
(BiTEs)
Bispescific
Two ScFvs
BiTEs form a link between T cells and
tumor cells
One of the scFvs binds to T cells via the
CD3 receptor, and the other to a tumor cell
via a tumor specific molecule
Blinatumomab (MT103): for the treatment
of non-Hodgkin’s lymphoma and acute
lymphoblastic leukemia; directed towards
CD19, a surface molecule expressed on B
cells.

41. Trifunctional antibody
Has 3 binding sites : Intact Fc , One binding
site for CD3, one for tumor antigen
Catumaxomab : (Fc, CD3, EpCAM)
The drug is approved (EMA) for the
treatment of malignant ascites in patients
with EpCAM-positive cancer if a standard
therapy is not available

42. Regulations

43. Competent Authorities
1. Review Committee on Genetic Manipulation (RCGM)
RCGM functions in the Department of Biotechnology (DBT):
responsible for authorizing import/export for research and
development and review of data up to preclinical evaluation.
2. Genetic Engineering Appraisal Committee (GEAC):
review and approval of activities involving large scale use of
genetically engineered organisms and products there of in research
and development, industrial production environmental release and
field applications.

44. 3. CDSCO:
responsible for grant of import/export license, clinical trial approval
and permission for marketing and manufacturing. State Food and
Drug Administration (FDA) works with CDSCO in each state and is
responsible for issuance of license to manufacture similar biologics in
India.

45. Scope
Guidelines address the regulatory pathway regarding
manufacturing process and quality aspects for similar biologics.
It also address the pre-market regulatory requirements including
comparability exercise for quality, preclinical and clinical studies
and post market regulatory requirements for similar biologics.
Any product can be considered as similar biologic only if it is
proven to be similar using extensive quality characterization
against the reference biologic.

46. These guidelines apply to similar biologics that contain well
characterized proteins as their active substance, derived through
modern biotechnological methods such as use of recombinant
DNA technology.
Similar biologic can only be developed against an authorized
reference biologic that has been approved using a complete data
package in India.
In case the reference biologic is not authorized in India, it should
have been licensed and marketed for at least 4 years with
significant safety and efficacy data.

47. In case of no medicine or only palliative therapy is available or
in national healthcare emergency, this period of 4 years may be
reduced or waived off.
The guidelines are applicable for similar biologics developed in
India or imported into the country.

48. Conclusion…
Monoclonal antibodies represent the largest and fastest growing type
of biopharmaceuticals.
Advances in genetic engineering over the years have provided
numerous ways to design MAbs that are more robust and efficacious
compared with their original murine version.
Their commercial and clinical success has fueled research activities
aiming to improve safety and efficacy.
Therapeutic antibodies have made the transition from conception to
clinical reality over the past two decades
In future, the information drawn from genomemedical science and
genome-informatics, that list the disease-related antigens useful for
medical treatment, should be essential to develop the therapy using
mAbs.

49. References
Katzung's - Basic and Clinical Pharmacology 12th edition
http://www.biologydiscussion.com/biotechnology/applicationsof-
monoclonalantibodies4applications/10045
http://www.reportlinker.com/p03312068summary/Advancesin-
MonoclonalAntibodyTherapeutics.html
Ansari W, Ghosh S. Monoclonal Antibodies: A tool in clinical research.
Ghosh. Indian Journal of Clinical Medicine 2013:4
Saeed AFUH, Awan SA (2016) Advances in Monoclonal Antibodies
Production and Cancer Therapy. MOJ Immunol 3(4): 00099. DOI:
10.15406/moji.2016.03.00099

50. Thank You !
Have a nice day !